黑水虻抗菌肽HI-3对RAW264.7细胞免疫调控作用研究

Immunomodulatory effect of antimicrobial peptide HI-3 from Hermetia illucens L.on RAW264.7 cells

为了明确黑水虻Hermetia illucensL.抗菌肽HI-3对RAW264.7细胞的免疫调控作用,拓展黑水虻抗菌肽在免疫领域的应用潜能,本文研究了抗菌肽HI-3对RAW264.7细胞形态、增殖、吞噬功能、分泌NO和细胞因子以及抗氧化活性的影响。结果表明,20、40、80、160、320μg/mL的抗菌肽HI-3对RAW264.7细胞形态及增殖指数均无显著影响,但均能显著增强RAW264.7细胞的吞噬功能,且随着HI-3浓度增加吞噬能力逐渐增强,当HI-3浓度达到320μg/mL时RAW264.7细胞的吞噬能力最强(P<0.01);而在160μg/mL以下处理浓度时,对NO含量无显著性影响(P>0.05);抗菌肽HI-3可以上调RAW264.7细胞分泌IL-6、TNF-α和IL-1β,下调抑炎因子IL-10的分泌量,且具有一定的浓度依赖性,浓度达到320μg/mL时效果最为显著(P<0.01);另外,抗菌肽HI-3亦能够显著提高RAW264.7细胞超氧化物歧化酶(SOD)活力和总抗氧化能力(T-AOC)含量,尤其是320μg/mL时,效果最为明显(P<0.01)。此外,320μg/mL以下处理浓度时对该细胞MDA含量影响不显著(P>0.05)。综上所述,黑水虻抗菌肽HI-3对RAW264.7细胞具有一定的免疫调控作用,具有良好的开发潜能。

In order to clarify the immunoregulatory effect of Hermetia illucens L.antimicrobial peptide HI-3 on RAW264.7 cells,and to expand the application potential of the antimicrobial peptide of H.illucens in the field of immunity.This article studied the effects of antimicrobial peptide HI-3 on the morphology,proliferation,phagocytosis,nitric oxide(NO),cytokines and antioxidant activity of RAW264.7 cells.The results showed that the antimicrobial peptide HI-3 at concentrations of 20,40,80,160 and 320μg/mL had no significant effect on the morphology and proliferation index of RAW264.7 cells,but both could significantly enhance the phagocytic function of RAW264.7 cells,and the phagocytic capacity gradually increased with the increase of HI-3 concentration.When the HI-3 concentration reached 320μg/mL,the phagocytic capacity of RAW264.7 cells was the strongest(P<0.01);When HI-3 was below 160μg/mL,it had no significant effect on NO content(P>0.05);The antimicrobial peptide HI-3 could also up-regulate the secretion of IL-6,TNF-α and IL-1β in RAW264.7 cells,and down-regulate the secretion of the anti-inflammatory factor IL-10,and it had a certain concentration dependence.The effect was most significant when the concentration reached 320μg/mL(P<0.01);In addition,the antimicrobial peptide HI-3 could also significantly increase the Superoxide dismutase(SOD)activity and Total antioxidant capacity(T-AOC)concentration of RAW264.7 cells,especially when 320μg/mL was the most effective(P<0.01).In addition,when the treatment concentration was below 320μg/mL the MDA content of the cell was also not significant(P>0.05).In conclusion,H.illucens antimicrobial peptide HI-3 had a certain immune regulation effect on RAW264.7 cells and showed good development potential.